SDS-PAGE Analysis of S. aureus Toxins

S. aureus CSs were analyzed by SDS-PAGE (4–20% gradient gel, BioRad, #456-1096) under reducing conditions. Proteins were transferred to PVDF (Hla, LukG and LukS-PV) or nitrocellulose membranes (LukD, HlgB) (Trans-Blot Turbo device, BioRad). Hla, LukD, and HlgB were detected using monospecific human IgGs, LukS-PV with a monoclonal mouse antibody (IBT Bioservices) and LukG expression using a rabbit anti-LukB (LukG) polyclonal antibody (IBT Bioservices) as described previously^{28 (link)}. Recombinant toxins expressed in E. coli^{28 (link)} were used as positive controls.

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Stulik L., Rouha H., Labrousse D., Visram Z.C., Badarau A., Maierhofer B., Groß K., Weber S., Kramarić M.D., Glojnarić I., Nagy G., Croisier D, & Nagy E. (2019). Preventing lung pathology and mortality in rabbit Staphylococcus aureus pneumonia models with cytotoxin-neutralizing monoclonal IgGs penetrating the epithelial lining fluid. Scientific Reports, 9, 5339.

Publication 2019

Trans-Blot Turbo device

Antibody Device Human Membranes Monoclonal antibody Mouse Nitrocellulose Proteins Pvdf Rabbit S aureus Sds page Toxins

Corresponding Organization :

Other organizations : Pliva (Croatia)

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Variable analysis

independent variables

Reduction conditions

dependent variables

Expression of Hla, LukD, LukG, LukS-PV, and HlgB proteins

control variables

SDS-PAGE (4-20% gradient gel, BioRad, #456-1096)
Transfer of proteins to PVDF (Hla, LukG and LukS-PV) or nitrocellulose membranes (LukD, HlgB) using Trans-Blot Turbo device (BioRad)
Detection methods: monospecific human IgGs (Hla, LukD, HlgB), monoclonal mouse antibody (LukS-PV), and rabbit anti-LukB (LukG) polyclonal antibody

positive controls

Recombinant toxins expressed in E. coli

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